TWI381845B - Composition for the prevention and treatment of Helicobacter pylori infection - Google Patents

Description

Composition for the prevention and treatment of Helicobacter pylori infection

The present invention relates to a composition for preventing and treating Helicobacter pylori infection in a non-antibiotic manner, and more particularly to a composition comprising catechin and sialic acid for preventing and treating Helicobacter pylori infection.

Helicobacter pylori is often associated with chronic active B gastritis, peptic ulcers, gastric cancer, and gastric mucosa-associated lymphoid tissue lymphoma. related. Currently, infection with H. pylori can be treated with a one-week treatment combined with a proton pump inhibitor and antibiotics. However, due to the increased resistance of H. pylori to some of the aforementioned drugs, the treatment of some patients is not effective. After initial treatment failure, second-line therapy, including triple or quadruple combination therapy, is usually recommended.

The first step in Helicobacter pylori infection is to penetrate the mucin and attach it to gastric epithelial cells through some different adhesion molecules. In vitro, anti-adhesive therapy often uses 3'-sialyllactose, which is an oligosaccharide-type sialic acid to prevent H. pylori binding. In the gastrointestinal epithelial cells, and in the animal experiments of rhesus monkeys, the colonization of H. pylori can be reduced without side effects. Usually, after H. pylori is attached to the gastric mucosa, gastric epithelial cells are damaged via Oxidative Stress, Apoptosis Type I or autophagy type II programmed death pathway.

Catechins belong to the group of Flavanol in polyphenols, also known as tea tannins, catechins, which are the general term for the flavanols in tea. The catechins are more tea. The most important one of the phenols accounts for about 75% to 80% of the tea polyphenol content. The derivatives of catechins are mainly divided into four types: epicatechin EC, epigallocatechin EGC, epicatechin gallate ECG, and table eclipse. Epigallocatechin gallate EGCG. Catechin and its derivatives Epigallocatechin gallate (EGCG) has antioxidant, anti-inflammatory, anti-apoptotic and cancer-preventing effects. In addition, catechins and EGCG are more resistant to food-borne pathogens, and by inhibiting Helicobacter pylori urease and vacuolating cytotoxin A (VacA), cytotoxin A (cytotoxin A) To achieve anti-Helicobacter pylori effect.

In today's antibiotic therapy for Helicobacter pylori, although it is effective most of the time, it sometimes fails due to the relationship between the resistance of the bacteria and the low compliance of the medication. Therefore, the use of non-antibiotics for the treatment of H. pylori infection has its urgent need.

Some strains such as Lactobacillus and Bifidobacterium have the effect of inhibiting H. pylori. However, it is known from clinical trials that the treatment of probiotics does not completely eradicate H. pylori. . In addition, the vaccine can also be administered prophylactically or therapeutically to the infection against H. pylori, but the results of the related mouse experiments show that although the amount of H. pylori can be significantly reduced, it cannot be completely eradicated. The effect of preventing infection is also limited.

There are certain non-antibiotic components that inhibit the growth of H. pylori, such as 3'-sialyllactose, an antioxidant molecule such as green tea catechin, which is an adhesion receptor antagonist. (tea catechins) has more credible efficacy. However, this effect only ends in the case of in vitro, and in the case of in vivo, the use of sialyllose or catechin alone cannot completely control the effect of inhibiting the growth of H. pylori.

These studies on the role of catechins and 3'-sialyl lactose inhibition in the development of Helicobacter pylori are only the result of individual use. Currently, catechin and 3'-sialyl lactose are combined to treat H. pylori infection. The effect is still unknown.

Due to the currently used drugs and methods, the problem of increased resistance of H. pylori cannot be solved. Therefore, in addition to the treatment of Helicobacter pylori infection by antibiotics, there is an urgent need for a composition which can reduce the infection of H. pylori or a combination of both preventive and therapeutic effects.

Accordingly, it is an object of the present invention to provide a composition for preventing H. pylori infection comprising: at least 0.128 wt% of one of catechin or a derivative thereof; at least 0.032 wt of one of sialic acid (silialic) Acid); and water. The weight percentage of each component is based on the total weight of the composition.

Another object of the present invention is to provide a composition for treating H. pylori infection comprising: 0.128 wt ‰ to 0.64 wt ‰ one of catechin or a derivative thereof; and one of 0.032 wt ‰ to 0.16 wt ‰ Silialic acid; and water. The weight percentage of each component is based on the total weight of the composition. To interfere with and reduce or eliminate H. pylori infection, that is, to block the action of Helicobacter pylori and gastric epithelial cells, while killing bacteria but not affecting gastric epithelial cells.

The invention adopts the technical means for solving the problem of reducing H. pylori infection, and adopts anti-adhesion components, anti-bacteria components and antioxidants, such as sialic acid (such as sialic acid). Sialic acid and the green tea extract, catechin or its derivatives, can produce protective functions against gastric damage induced by Helicobacter pylori. This effect is mainly through reducing the amount of O ₂ ^- , H ₂ O ₂ , NO produced in cells and the expression of inducible nitric oxide synthase (iNOS), increasing the antioxidant activity of cells, and through autophagy ( Autophagy) and bidirectional regulation of apoptosis (apoptosis). Especially after combining sialic acid and green tea extract-catechin, it can up-regulate the path of autophagy that protects the living cells and reduce the damage of gastric damage induced by Helicobacter pylori. Through a series of in vivo and in vitro experiments, it is confirmed that the use of catechin and sialic acid can prevent and treat Helicobacter pylori infection.

Through the technical means adopted by the invention, the Helicobacter pylori can effectively reduce the injury of gastric epithelial cells and alleviate the phenomenon of gastric inflammation, and can achieve complete inhibition of up to 100% in prevention, and can reach 60% in treatment. Inhibitory effect. Both of these main ingredients can be obtained from natural foods, and can be obtained through a simple pipeline and ingested foods rich in sialic acid and green tea extracts to prevent or treat symptoms caused by Helicobacter pylori infection in the stomach.

One of the ingredients of the present invention is catechin or a derivative thereof. The catechin or its derivative is epigallocatechin gallate (EGCG), epicatechin gallate (ECG), and a gallic catechin. Acidic acid gallate (Gallocatechin gallate GCG), an epicatechin EC, epigallocatechin EGC, gallocatechin GC, and any combination thereof.

The "Sialic acid" used in the present invention is a naturally occurring 9-carbon monosaccharide derivative known by the name "N-acetylthioneine", which can be used as a negatively charged ion to make saliva. Produces a smooth feel.

The embodiments of the present invention are further described in the following description, and the embodiments of the present invention are set forth to illustrate the present invention, and are not intended to limit the scope of the present invention. In the scope of the invention, the scope of protection of the invention is defined by the scope of the appended claims.

The statistical analysis used in the examples of the present invention, here using the score test and the binomial test, showed significant differences in the effect of combining catechin/sialic acid against Helicobacter pylori in each group. Sex. In cell culture experiments, one-way ANOVA and Duncan's Multiple Range test were used to characterize the significance of differences between groups in independent samples between groups. In the animal model experiment, the Kruskal-Wallis test, One-way ANOVA and Dunnett's multiple comparison were used to verify each. The significance of the difference between the groups in the independent samples. Logistic Regression was applied to show the dose effect in terms of antimicrobial rate. P-value <0.05 is considered to be statistically significant.

The embodiments of the present invention are exemplified for convenience of description, but are not limited to the invention of the present invention. In the following examples, all amounts, parts, or percentages are based on total weight or total amount unless otherwise stated.

Strains and drugs

The H. pylori used in the examples includes the standard strain ATCC 43504 and 20 clinical isolates (TA1-TA20). The clinical isolates used herein were obtained from gastric sections of patients with gastritis or peptic ulcer.

The decaffeinated green tea extract was purchased from Vigour Biochemistry, including 328 mg/g epigallocatechin gallate EGCG and 152 mg/g epicatechin gallic acid ( Epicatechin gallate ECG), 148 mg/g gallocatechin gallate GCG, 132 mg/g epicatechin EC, 108 mg/g epigal tea Epigallocatechin EGC, 104 mg/g of epicatechin GC and 44 mg/g of catechin. The sialic acid was purchased from Sigma.

Embodiment 1 In vitro anti-Helicobacter pylori activity of catechins and sialic acid

First, an in vitro test of catechin and sialic acid against various Helicobacter pylori strains against H. pylori was performed. The 20 strains to be tested were stored at -80 ° C and recovered in a micro-aerobic environment (5% O ₂ , 10% CO ₂ , 85% N ₂ ) at 37 ° C for 3 days, then 10 mL. The Brucella broth was resuspended for 24 hours until it reached an optical density of 0.5 unit at 450 nm (corresponding to about 10 ⁹ CFU/L). The minimum inhibitory concentration (MIC) of catechin and sialic acid is determined by the Agar dilution method. The effect of combining catechin and sialic acid was determined by the checkerboard method and evaluated by fractional inhibitory concentration index (FIC).

The results are shown in Table 1, which shows the results of in vitro tests of catechin and sialic acid against the anti-Helicobacter pylori effect of 20 Helicobacter pylori strains. The method for obtaining the minimum inhibitory concentration (MIC) is as described above. The results are shown in Table 1. The minimum inhibitory concentration (MIC ₉₀ ) of catechin for 90% of the isolated strains was 256 mg/L. From the results, it can be seen that the use of sialic acid alone does not have any anti-Helicobacter pylori effect.

Embodiment 2 In vitro anti-Helicobacter pylori activity of catechin and sialic acid

Next, catechin and sialic acid-binding in vitro anti-Helicobacter pylori activity tests were also carried out using 20 strains. The results are shown in Table 2, which shows the antibacterial effect of the combined use of catechin and sialic acid on the Helicobacter pylori strain. The method for determining the score inhibition concentration index (FIC) is as described above. As shown in Table 2, the simultaneous use of catechin and sialic acid has an effect on all clinical isolates of Helicobacter pylori, with an additive effect or a synergistic effect, respectively. It can be seen from the results that sialic acid enhances the antibacterial effect of catechin. In addition, as a result of the checkerboard method (not shown), in vitro experiments with 128mg/L catechin and 32mg/L sialic acid can completely inhibit the growth of all clinical isolates, whether antibiotic sensitive or resistant The strains of the type were all the same.

Embodiment 3 Human gastric cancer cells (AGS cells) infected with Helicobacter pylori, reduced oxidative stress after treatment with catechin and sialic acid Cell culture method

Using a Cytotoxin-Associated Gene A-/Vacuolating cytotoxin A-positive H. pylori strain (TA1) in a Colombian agar plate containing 5% sheep blood It was recovered from the frozen state (-80 ° C) at a temperature of 37 ° C for 3 days in a micro aerobic environment. Human gastric cancer cells ATCC CRL 1739 (AGS cells) were supplemented with 10% fetal bovine serum (FBS) in RPMI 1640 medium (Invitrogen). In order to simultaneously culture H. pylori and AGS cells, the cells were resuspended in PBS to an OD ₄₅₀ of 1 unit, corresponding to a bacterial concentration of about 2 x 10 ¹¹ CFU/L, and the ratio of H. pylori to AGS cells was added. The cells were co-cultured for 4 hours in a 100:1 well plate with or without 128 mg/L catechin and/or 32 mg/L sialic acid.

2.NO, O ₂ ^- And H ₂ O ₂ Quantification of concentration

The measurement of the concentration of nitric oxide (NO) was measured by a NO chemical luminescence probe and a chemical luminescence analysis system (CLD-110, Tohoku Electronic). On the other hand, in order to measure the concentration of O ₂ ^- and H ₂ O ₂ , chemistry was carried out by adding 0.2 mL of the culture sample and a sensitizing agent (0.1 mmol/L of lucigenin or 0.2 mmol/L of luminol) to PBS (pH 7.4). In the cold light method, the test was performed in three repetitions and the chemical luminescence intensity was counted every 10 seconds.

3. Quantification of iNOS performance

Using Western blotting method, cells treated under different conditions for 4 hours, extracted their proteins, analyzed by SDS-PAGE electrophoresis on a 10% polyacrylamide gel, and transferred to a semi-dry transfer system (Hoeffer Phamacia Biotech). It was printed on a PVDF (polyvinylidine difluoride) membrane, and the PVDF membrane was immersed in PBS containing 5% milk for 2 hours (at room temperature), and then placed in an antibody containing anti-iNOS (inducible NO synthase, Chemicon) and anti-Bax. Soaking solution of antibody, anti-Bcl-2 antibody, anti-caspase 3 antibody, anti-poly-(ADP-ribose)-polymerase antibody (all purchased from cell signaling Technology) or anti-Beclin-1 (BD Biosciences) antibody After washing for 3 times with a washing solution for 1 hour, it was treated with a blocking buffer containing HRP-labeled goat anti-rabbit immunoglobulin IgG antibody (Pierce) for 1 hour, and developed with ECL solution (Amersham Biosciences), and Exposure to X-ray film.

Referring to Figures 1A to 1D, O ₂ ^- , H ₂ O ₂ , in the case of AGS cells infected with Helicobacter pylori (HP) for 4 hours, using catechin (C) and sialic acid (S), The amount of NO and iNOS produced varies, and the 1D map also includes the results of the Western blot method. It can be seen from previous studies that infection with H. pylori can cause DNA damage and increase the production of active oxides (ROS) and the expression of inducible nitric oxide synthase (iNOS), thereby observing the antioxidant activity in cells. As a result, as shown in Fig. 1, after infection with Helicobacter pylori (HP), the production of O ₂ ^- , H ₂ O ₂ and NO increased, and the expression of iNOS also increased. This effect lasts from about 1 hour after infection with H. pylori to about 4 hours, and when catechin and sialic acid are used simultaneously, the amount of active oxide (ROS) is increased and inducible nitric oxide synthase ( The situation in which iNOS) increased in performance was significantly suppressed. The above results show that the use of catechin (C) and sialic acid (S) promotes the antioxidant action of cells.

Embodiment 4 Human gastric cancer cells (AGS cells) infected with Helicobacter pylori, decreased apoptosis of AGS cells treated with catechin and sialic acid

Refer to Figures 2A to 2B, which show the expression of apoptosis-related proteins in the case of AGS cells infected with Helicobacter pylori (HP) for 4 hours, using catechin (C) and sialic acid (S). The electrophoresis analysis conditions are as described above. Among them, Bax, Bcl-2, CPP32 and PARP are proteins related to apoptosis, and the ratio of both Bax and Bcl-2 is mainly observed as an index. As shown in Figures 2A and 2B, AGS cells increased the expression of Bax after infection with H. pylori, while the expression of Bcl-2 decreased, indicating that the ratio of Bax:Bcl-2 increased after infection with H. pylori, making AGS The situation in which cells undergo apoptosis is increased. After the use of catechin and sialic acid, the performance of Bax in infected AGS cells was decreased, and the expression of Bcl-2 was increased, indicating that AGS cells can be apoptotic after using catechin and sialic acid. cut back. In addition, the expression levels of CPP32 (i.e., caspase 3) and PARP (i.e., poly-(ADP-ribose)-polymerase) of infected AGS cells were significantly inhibited after the simultaneous use of catechin and sialic acid. It showed a decrease in apoptosis of AGS cells after catechin and sialic acid treatment.

Embodiment 5 Human gastric cancer cells (AGS cells) infected with Helicobacter pylori, increased autophagy in AGS cells treated with catechin and sialic acid

Referring to Fig. 3, it is shown that the amount of autophagy-related protein Beclin-1 is changed in the case where AGS cells are infected with Helicobacter pylori (HP) for 4 hours, using catechin (C) and sialic acid (S). In the case, the electrophoresis analysis conditions are as described above. Among them, Beclin-1 is a Bcl-2 interacting protein, which plays a role in promoting autophagy. As shown in the figure, AGS cells reduced the expression of Beclin-1 after infection with H. pylori, but the expression of Beclin-1 increased after the use of catechin and sialic acid, indicating that it promoted Beclin-1 dependence. Autophagy (Beclin-1-dependent autophagy).

Next, changes in the cell type of catechin (C) and sialic acid (S) were observed after 4 hours of infection with H. pylori (HP) by AGS cells by a fluorescence microscope. Autophagic vacuoles were calibrated with a triple repeat of 0.05 mmol/L monodansylcadaverine (MDC). After the calibration, it was washed 4 times with PBS and immediately fixed with 4% paraformaldehyde and placed under a fluorescent microscope (Leica model DMRD). The apoptosis of AGS cells induced by H. pylori was evaluated by terminal deoxynucleotidyl transcferase mediated nick end labeling.

The results are shown in Figures 4A to 4L. It shows that AGS cells undergo type change, apoptosis and catechin (C) and sialic acid (S) after 4 hours of infection with Helicobacter pylori (HP). In the case of autophagy, the 4A~4D images are images observed by phase contrast microscopy in the absence of staining, and the 4E~4H images are stained by the terminal deoxynucleotidyl transferase-mediated nick end labeling. The 4I~4L images were stained with 0.05 nM MDC.

The 4A, 4E, and 4I maps are the type of intact AGS cells, the absence of apoptosis, and the phenomenon of mild autophagy. As shown in Figures 4B, 4F, and 4J, changes in morphology, formation of apoptosis, and inhibition of autophagy are observed after infection with H. pylori. If combined with catechin and sialic acid treatment, it can maintain the shape of AGS cells, inhibit apoptosis, and maintain autophagy, as shown in 4C, 4G and 4K. In the cells that are not infected with H. pylori, the combination of catechin and sialic acid does not damage the cells, as shown in Figures 4D, 4H and 4L.

Embodiment 6 Animal model and result detection

The 5-week-old BALB/c mice (provided from the National Experimental Animal Center) were grown at a constant temperature in the National Taiwan University Animal Experimental Center. Mouse feed (20.5% protein, 18.5% fat, 53% carbohydrate, 2.7% cellulose, 4.8% mineral) and any amount of drinking water. All of the procedures and experimental procedures were approved by the Animal Humanitarian Care Committee of the National Taiwan University and in accordance with the instructions of the National Science Council.

Forty mice infected with H. pylori were divided into 4 groups in groups of 10 each. TA1 was used as an infected mouse. The colonies were transferred to Brucella broth plus 5% fetal bovine serum, 1% VitaleX and antibiotics (48 hours), after which the bacterial concentration was adjusted to 10 ¹¹ CFU/L. Each group of mice was orally infused with 0.5 mL of bacterial solution for two consecutive days. In the control group, distilled water was injected. Mice that require pretreatment are given 0.5 ml of distilled water containing 128 mg/L catechin and 32 mg/L sialic acid (72 hours ago) before injecting H. pylori into the mouth, and then freely ingesting 128 mg. / L catechin and 32 mg / L of sialic acid 1% glucose water (for 3 consecutive days). The post-treated group was fed with 0.5 ml of distilled water containing 128 mg/L catechin and 32 mg/L sialic acid two weeks after infection (after 2 weeks of infection with H. pylori), and then freely ingested with 128 mg. / L catechin and 32 mg / L of sialic acid 1% glucose water (for 5 consecutive days). The control group infected with Helicobacter pylori was orally administered with 1% glucose water from 3 days before infection to 5 days after infection. Except for the steps described above, the other steps are the same. Four weeks after H. pylori infection, the stomach was examined and cut into two equal portions for histological and microbiological tests.

The tissue staining method uses 3-nitrotyrosine 3-NT and 4-hydroxynonenal 4-HNE to display in situ in the H. pylori-infected tissue, that is, Immunostaining of paraffin-embedded parts was performed by 3-NT and 4-HNE. Rabbit anti-nitrotyrosine IgG antibody (NITT12-A) or rabbit anti-nitrotyrosine antibody (NNE11-S) was allowed to stand overnight at 4 °C (both purchased from Alpha Diagnostics at 1:50) Dilute in PBS). Staining was performed in a kit of avidin-peroxidase complex method (ABC Elite, Vector Laboratories). The stained site was visualized with diaminobenzidine tetrahydrochloride and counterstained with Hematoxylin.

After 3-5 days of culture, it is possible to confirm whether or not H. pylori infection is caused by the above-mentioned tissue staining and microbiological test results, the degree of gastritis is distinguished by a pathologist, and the number of cells can be calculated after microbiology culture. (CFU/L). In addition, the presence of H. pylori can be confirmed by PCR.

Referring to Table 3, it was shown that BALB/c mice infected with H. pylori had the effect of combining catechin and sialic acid. From the results in the table, all the infection control groups of H. pylori were successfully infected, and the gastric mucosa had significant edema and hemorrhage.

At the same time, refer to the 5A~5N figure, which shows the pathological section analysis of the stomach of BALB/c mice after infection with Helicobacter pylori. Observed by microscopy, significant gastritis can be seen, accompanied by many monocytes and neutrophil infiltration phenomena, as shown in Figures 5F-5H. Figure 5F shows inflammation of the gastric mucosa and submucosa (H&E staining, 100 times), Figure 5G shows gastric mucosal destruction after infection (H&E staining, 400 times), and Figure 5H shows multiple infections after infection. Sexual neutrophil aggregation and microabscess (H&E staining, 400 times). The average gastritis score is 2.0 (see Table 3).

In the group pretreated with catechin and sialic acid, no gastric mucosal lesions were observed due to infection with H. pylori, and only minimal tissue changes were observed from the section results. Fig. 5C~5D), and the average gastritis score is 0.3 (see Table 3), which is the same as the group that only takes distilled water and is not infected with H. pylori (see Figures 5A-5B).

Fig. 5E shows the phenomenon of post-treatment of Helicobacter pylori (H&E staining, 100-fold) after post-treatment with catechin and sialic acid. About 20% of mice have been infected with H. pylori infection. Although most of the mice that did not eliminate H. pylori infection still had gastric mucosal lesions, the average gastritis score was reduced to 0.8 (compared to the infected group). The uninfected, pretreated, and posttreated groups were significantly different from the infected control group (p < 0.01 for each group).

Refer to the 5I~5N map, the 5I~5K picture shows the color reaction of oxidative 3-NT staining (brown) in the proximal gastric mucosa, wherein the 5I picture is the uninfected control group, and the 5th picture is the infected group, The 5K picture is the pre-processing group. The 5L~5N picture shows the color reaction of oxidized 4-HNE staining (brown) in the proximal gastric mucosa, the 5th picture shows the uninfected control group, the 5th picture shows the infected group, and the 5th picture shows the pretreatment group. First, from the results in the 5I~5K chart, the uninfected control group (see Figure 5I) is blue without oxidation, and infected with Helicobacter pylori (see Figure 5J) is obviously brown, with catechin and sialic acid. The group that did the pre-infection treatment (see Figure 5K) was also blue without oxidation. Next, from the results of the 5L~5N map, the control group (see Figure 5L) is blue without oxidation, and infected with Helicobacter pylori (see Figure 5M) is obviously brown, and is made with catechin and sialic acid. The pre-infection treatment group was blue without oxidation (see Figure 5N). Briefly, 3-NT and 4-HNE-accumulated adducts were observed proximal to the stomach of infected mice, and their accumulation was much higher than that pretreated with catechin and sialic acid. Group.

Post-treatment with catechin and sialic acid inhibits H. pylori infection in a dose-dependent manner

The same procedure as described above for post-treatment was used to test the effect of different concentrations of catechin/sialic acid in the treatment of H. pylori infection. Sixty mice were divided into three groups for post-treatment [at the same dose (1 STD) or 2, 5-fold dose (2, 5 STD)]. Another 10 mice served as control group for infection (none-treatment; NT). The cure rate for each group was evaluated 4 weeks after treatment with catechin/sialic acid.

The eradication rate of H. pylori in the group treated with catechin and sialic acid after infection was 0%, treated with 1 standard dose (128 mg/L catechin and 32 mg/L sialic acid) The eradication rate of H. pylori in the group was 20%, the eradication rate of H. pylori in the group treated with 2 times the dose was 30%, and the eradication rate of H. pylori in the group treated at 5 times was 60%. (as shown in Figure 6). This dose-dependent effect on eradication ratio was statistically significant (p < 0.01), and the Odds Ratio (OR) = 1.695 for each additional standard dose.

It can be seen from the foregoing description and the examples that the catechin and sialic acid compositions of the present invention can achieve 100% complete inhibition of Helicobacter pylori infection and up to 60% eradication rate after infection treatment. It is proved that the composition containing catechin and sialic acid has a remarkable effect on anti-Helicobacter pylori. The foregoing methods are merely examples and are not intended to limit the scope of the invention.

Figures 1A to 1D show O ₂ ^- , H ₂ O ₂ in the case of AGS cells using catechin (C) and sialic acid (S) 4 hours after infection or uninfected with Helicobacter pylori (HP), respectively. , NO and iNOS changes;

Figures 2A to 2B show the expression of apoptosis-related proteins in the case of AGS cells using catechin (C) and sialic acid (S) 4 hours after infection or uninfected with Helicobacter pylori (HP). ;

Figure 3 shows the expression of autophagy-associated protein Beclin-1 in the case of AGS cells treated with catechin (C) and sialic acid (S) 4 hours after infection or uninfected with Helicobacter pylori (HP). ;

The 4A~4L map shows the type change, apoptosis and the use of catechin (C) and sialic acid (S) after 4 hours of infection or uninfection of Helicobacter pylori (HP). The situation of autophagy in cells;

Figures 5A-5N show the pathological section analysis of the stomach of BALB/c mice after infection or not infection with Helicobacter pylori.

Figure 6 shows the clearance rates of different dose groups using different doses of catechin and sialic acid in infected BALB/c mice.

Claims (14)

A composition for preventing infection by H. pylori, comprising: at least 0.128 wt One of catechins or derivatives thereof; at least 0.032 wt a sialic acid; and water, wherein the Helicobacter pylori strain comprises a strain resistant to metronidazole or clarithromycin, and the catechin and its derivative are combined with sialic acid At the same time, oral administration was continued for at least 3 days to eradicate H. pylori infection. The composition for preventing Helicobacter pylori infection according to Item 1, wherein the catechin or a derivative thereof is selected from the group consisting of epigallocatechin gallate (EGCG), Epicatechin gallate ECG, Gallocatechin gallate GCG, epicatechin EC, a table of tea Epigallocatechin EGC, gallocatechin GC, and any combination thereof. The composition for preventing Helicobacter pylori infection according to the above item 1, wherein the catechin is extracted from a tea extract. The composition for preventing Helicobacter pylori infection according to claim 3, wherein the tea extract is a decaffeinated green tea extract. The composition for preventing Helicobacter pylori infection according to the above item 1, wherein the composition is applied to a stomach. The composition for preventing Helicobacter pylori infection according to Item 1, wherein the composition reduces the production of O ₂ ^- , H ₂ O ₂ , NO, and inducible nitric oxide synthase (iNOS) in the cell. The amount of expression to increase the antioxidant activity of normal cells. A composition for treating H. pylori infection, comprising: 0.128 wt To 0.64 wt One of catechins or derivatives thereof; 0.032 wt To 0.16 wt a sialic acid; and water, wherein the Helicobacter pylori strain comprises a strain resistant to metronidazole or clarithromycin, and the catechin and its derivative are combined with sialic acid At the same time, oral administration was continued for at least 2 weeks to eradicate H. pylori infection. The composition for treating H. pylori infection according to claim 7, wherein the catechin or a derivative thereof is selected from the group consisting of epigallocatechin gallate (EGCG), Epicatechin gallate ECG, Gallocatechin gallate GCG, epicatechin EC, a table of tea Epigallocatechin EGC, gallocatechin GC, and any combination thereof. The composition for treating H. pylori infection according to claim 7, wherein the catechin is extracted from a tea extract. The composition for treating H. pylori infection according to claim 9, wherein the tea extract is a decaffeinated green tea extract. The composition for treating H. pylori infection according to claim 7, wherein the composition is applied to a stomach disease. The composition for treating Helicobacter pylori infection according to claim 11, wherein the gastric disease is chronic active B gastritis, peptic ulcers, gastric cancer, Or gastric mucosa-associated lymphoid tissue lymphoma. The composition for treating Helicobacter pylori infection according to claim 7, wherein the composition reduces the production of O ₂ ^- , H ₂ O ₂ , NO and inducible nitric oxide by reducing the amount of O ₂ ^- , H ₂ O ₂ , NO in an infected cell. The amount of enzyme (iNOS) is expressed to increase the antioxidant activity of the infected cells. The composition for treating H. pylori infection according to claim 7, wherein the composition protects the viable autophagy pathway by up-regulation and reduces apoptosis of gastric lesions induced by Helicobacter pylori (apoptosis) damage.